应用梭状芽孢杆菌口服疫苗载体表达HIV p24蛋白的研究

应用一种不产毒的梭状芽孢杆菌（Clostridium perfringens,C.P.）作为口服疫苗载体表达HIVp24蛋白。用PCR扩增HIVp24基因,酶切后插入到含有C.P.序列的pJRC200质粒的CPEORF中,将重组的cp24质粒转化无致病作用的ATCC3624菌株。使HIVp24基因在C.P.形成芽孢时大量表达。结果应用SDS_PAGE和West Blot分析HIVp24在重组C.P.繁殖体和芽孢中的表达情况,发现HIVp24蛋白在C.P.芽孢体中大量表达,而在C.P.繁殖体中却没有表达。证明在体外构建了含有HIVp24基因的重组pJRC200质粒,成功地表达了HIVp24蛋白并得到了HIVp24的高表达ATCC3624菌株。为成功研究C.P.作为一种口服HIV疫苗载体奠定了基础。

Use of a Clostridium perfringens as an Oral HIV Vaccine Vector to Express High Levels of HIV p24 Protein

Use of a noncytotoxic Clostridium perfringens as an oral HIV vaccine vector to express high levels of HIV p24 protein,the entire HIV p24 ORF was amplified by PCR, then digested and ligated with plasmid pJRC200 ,which contained a Clostridium perfringens DNA and had been removed most of the CPE ORF. The recombinant cp24 was transformed into a noncytotoxic Clostridium perfringens strain ATCC3624 (cpe-negative, type A) to be highly expressed during sporulation. It is developed a recombinant plasmid pJRC200, which contained HIV p24 ORF. HIV p24 ORF was expressed successfully. A highly expressed HIV p24 strain ATCC3624 was abstained. This work will be a base for the study of the use of Clostridium perfringens as an oral HIV vaccine.